Gel Electrophoresis (original) (raw)
Related papers
Quantitative analysis of the three regimes of DNA electrophoresis in agarose gels
Biopolymers, 1988
An extensive series of experiments has been performed to study the mobility of DNA fragments ranging in size from 2.0 to 48.5 kilobose pairs. By varying the agarose concentration in the gels and the electric field strength, three DNA electrophoresis regimes were clearly identified: the Ogston regime (small DNA fragments in large pores of agarose), the reptation regime without DNA chain stretching (small pores of agarose and weak electric fields), and the reptation regime with DNA chain stretching (small pores of agarose, strong electric fields, and large DNA fragments). Here we report on the experimental identification of these regimes and on the conditions governing the transition between each of them. The onset of reptation and of stretching of DNA chains in gel electrophoresis are described quantitatively for the first time, and a phase diagram for the dynamics of DNA during electrophoresis is presented.
Electrophoresis-staining apparatus for DNA agarose gels with solution exchange and image acquisition
Instrumentation Science & Technology, 2016
Agarose gel electrophoresis is routinely used for the separation of nucleic acids. Gels are developed, stained, and visualized using dedicated equipment and reagents. Manufacturers have developed instrumentation with advanced features that provide good safety and user-friendly operation. However, the process of size fractionation of nucleic acids by horizontal gel electrophoresis by dye staining may be cumbersome and unsafe due to many steps and harmful chemicals. Here is reported a safe, inexpensive, time-saving, and comprehensive apparatus for gel electrophoresis, staining, and imaging. This newly modified apparatus has simple operation and uses existing equipment and off-the-shelf components for easy construction in the laboratory. The apparatus has been shown to perform agarose horizontal gel electrophoresis and associated techniques with ease and simplicity.
Hoechst 33258 staining of DNA in agarose gel electrophoresis
Journal of Microbiological Methods, 1986
A staining procedure for agarose gel electrophoresis using Hoechst 33258 has been optimized. A concentration of 5 x 10-s M allows the immediate visualization of as little as 1 ng of DNA with a standard UV illumination system. The major advantage of this method is the elimination of the carcinogen ethidium bromide used in most DNA visualization methods for gel electrophoresis.
Electrophoresis, 1995
A comparison of resolution of DNA fragments between agarose gel and capillary zone electrophoresis in agarose solutions The resolving power of capillary zone electrophoresis (CZE) is compared to that of gel electrophoresis (GE) under similar conditions (agarose, similar length of DNA fragments, identical buffer) but with differences in temperature and field strength. The comparison is based on the time required to reach a desired degree of resolution by each of the two methods. A resolution parameter is developed which is equally applicable to CZE, with relatively diffuse initial conditions in the absence of stacking and measurements expressed in terms of time, and to GE, in which measurements are expressed in terms of spatial parameters. The resolution time in CZE using agarose solutions at 40°C was found to be greater by at least one order of magnitude than that in GE using agarose gels. Thus, the increased migration velocity due to high field strength in CZE substantially outweighs the lower dispersion in GE.
Separation and analysis of DNA by electrophoresis
Current Opinion in Biotechnology, 1991
Pulsed-field gel electrophoresis, the polymerase chain reaction and capillary electrophoresis represent major technical advances in DNA electrophoresis in the last few years. This has lead to a number of new applications, including the cloning of large DNA fragments, multiplex analysis of DNA samples, and the detection of single base changes in DNA. New fluorescent and chemiluminescent techniques promise to simplify detection methods. Current Opinion in Biotechnology 1991, 2:86-91 Abbreviations ADP-allele-dependent primer; ASO-allele-specific oligonucleotide; DGG[-denaturing gradient gel electrophoresis; FIG~ield inversion gel electrophoresis; PCR-polymerase chain reaction; PFG~pulsed-field gel electrophoresis; YAC-yeast artificial chromosome.
Biochemistry, 1988
The resolution of pulsed-field gel electrophoresis is dramatically affected by the number and configuration of the electrodes used, because these alter the shape of the applied electrical fields. Here we present calculations and experiments on the effect of electrode position in one of the most commonly used pulsed-field gel electrophoresis configurations. The goal was to explore which aspects of the electrical field shape correlate with improved electrophoretic resolution. The most critical variable appears to be the angle between the alternate electrical fields. The most effective electrode configurations yield angles of more than 1 loo. A continually increasing angle between the fields produces band sharpening that greatly enhances the resolution.